Integrative RNA-Seq Profiling Unveils Compensatory Transcriptional Networks in Prm1-knockout Evolved Strains of Schizosaccharomyces pombe

Comparative RNA-Seq profiling was conducted to elucidate the transcriptional adaptations that underlie the adaptive increase of cell fusion efficiency in Schizosaccharomyces pombe. The study systematically analyze three strain groups: (1) wild-type, (2) ancestor (engineered Prm1 knockout in wild-type background), and (3) serial prm1-knockout evolutionary strains derived through single-colony selection experimental evolution under successful fusion selection in cycle 3, 6, 9, 12, 15, 18. Evolved lineages showing restored fusion efficiency were prioritized for sequencing.RNA was extracted at 0 h and 5 h post nitrogen starvation in MSL-N medium to profile starvation-responsive pathways, with biological triplicates ensuring statistical robustness. Libraries prepared using stranded poly-A selection (NEBNext Ultra II) were sequenced on Illumina NovaSeq platform with paired-end reads (150 bp, 2G).